Navigation Links
Laser Capture Microdissection of muscle fiber populations and expression analysis by RT-PCR

◊ Horizontal gel chamber
◊ Power supply
◊ Gel Doc 2000TM Documentation System (Bio-Rad, Cat. # 170-8615)


RNase-free Technique
In addition to the usual precautions listed below, some special precautions were taken connected to the rapid staining protocol:
1. Use RNase AWAY according to the manufacturers instructions on laboratory bench surfaces, cryostat, cryostat knife, PixCell II Laser Capture Microdissection System and Arcturus alignment tray.
2. Disposable gloves are to be frequently changed and RNase-free plasticware used.
3. Heat inactivate serum.
4. Use DEPC-treated water for the preparation of all staining solutions and for washing.
5. Use chemicals in the highest grade available.
6. Staining and washing time is to be reduced to a minimum, while still preserving the capability to distinguish different muscle fiber populations.


Method
1. After dissection, limb muscle and EOMs are covered in OCT compound and successively frozen in cooled 2methylbutane (30 seconds) and liquid nitrogen (10 minutes). The muscle can be stored at -80C.

2. The muscle is cut into 10 m sections with the cryostat. During the cutting process, the slides with 4 sections each are stored on dry ice and are afterwards stored at -80C.

3. For the acetylcholine esterase stain by Karnowsky and Roots, the following solution is prepared in the following order:
5 mg of acetylthiocholine iodide are dissolved in 6.5 ml of 0.1 M sodium acetate buffer pH 5.2, followed by:
0.5 ml 0.2 M sodium citrate,
1.0 ml of 30 mM cupric sulfate,
1.0 ml DEPC-treated water, and
1.0 ml of 5 mM potassium ferricyanide.

Larger q
'"/>

Source:


Page: All 1 2 3 4 5 6 7 8

Related biology technology :

1. Isolate and Analyze Total RNA from Cells Harvested by Laser Capture Microdissection
2. Laser microdissection and nucleic acid purification - a Leica - QIAGEN cooperation
3. Working With Laser Capture Microdissection Samples
4. Laser Capture Microdissection (LCM) for the Analysis of Macrophage Gene Expression from Atherosclerotic Lesions
5. Laser Capture Microdissection (LCM) and Expression Profiling of Long-Range Projection Neurons
6. Differential Gene Expression Analysis of Pure Cell Populations from Frozen UterineTissue Using Laser Capture Microdissection (LCM) and cDNA Microarrays
7. Laser Capture Microdissection of Cells Labeled with Enhanced Green Fluorescent Protein
8. Primary cells, rat heart muscle
9. High-throughput gene expression analysis to screen for anti-asthma drugs
10. Quantitative end-point RT-PCR gene expression measurement using the Agilent 2100 Bioanalyzer and standardized RT-PCR
11. Cytosolic expression of Green Fluorescent Protein (GFP) and its derivatives in the yeast Saccharomyces cerevisiae: Detection in vivo using the Varian Cary Eclipse
Post Your Comments:
(Date:7/22/2014)... (PRWEB) July 22, 2014 Automated cell ... and maintenance of cell culture. They allow production of ... needs of researchers in drug discovery and industries and ... in mAb production, iPSC research, cryobanking, cell-based assays, and ... contamination risk. , Analysts forecast the Global Cell ...
(Date:7/22/2014)... Mich., July 22, 2014  Neogen Corporation (NASDAQ: ... 2014 fiscal year, which ended May 31, increased 19% ... income was $28,158,000, compared to the prior year,s $27,190,000. ... 2013, earnings per share in the current year were ... and net income for the 2014 fiscal year established ...
(Date:7/22/2014)... July 22, 2014 Hussey Copper , ... 29 antimicrobial copper announces Pullman Regional Hospital,s conversion ... EPA-registered antimicrobial solid touch surface, at a nominal cost.  ... MRSA, E-coli and other bacteria within 2 hours of exposure ... against bacteria and infectious diseases in its facilities. ...
(Date:7/22/2014)... , July 22, 2014  United Therapeutics Corporation ... it will release its second quarter 2014 financial ... 2014. United Therapeutics will host a ... 9:00 a.m. Eastern Time.  The teleconference is accessible ... A rebroadcast of the teleconference will be available ...
Breaking Biology Technology:Global Cell Culture Market to Grow at 10.27% CAGR by 2018 Forecasts a New Research Report ReportsnReports.com 2Global Cell Culture Market to Grow at 10.27% CAGR by 2018 Forecasts a New Research Report ReportsnReports.com 3Global Cell Culture Market to Grow at 10.27% CAGR by 2018 Forecasts a New Research Report ReportsnReports.com 4Neogen reports 19% increase in annual revenues 2Neogen reports 19% increase in annual revenues 3Neogen reports 19% increase in annual revenues 4Neogen reports 19% increase in annual revenues 5Neogen reports 19% increase in annual revenues 6Neogen reports 19% increase in annual revenues 7Neogen reports 19% increase in annual revenues 8Pullman Regional Hospital changes more than 1,100 touch points to Hussey Copper's MD-Cu29 antimicrobial copper to help protect against bacteria 2Pullman Regional Hospital changes more than 1,100 touch points to Hussey Copper's MD-Cu29 antimicrobial copper to help protect against bacteria 3
... SAN FRANCISCO, May 10, 2011 Date: ... ranking the 500 fastest growing technology, media, telecommunications, life sciences, and ... of the Technology Fast 500™. What:   Nominations ... revenue growth over fiscal years 2006 to 2010.  To be eligible ...
... DATATRAK International, Inc. (OTCQX: DATA), a technology and ... clinical trials industry, today announces the expansion of its ... Community College (DTCC) to license its DATATRAK eClinical® software ... software in its Clinical Trials Research program and courses. ...
... COLLEGE PARK, Md., May 9, 2011 The Maryland ... the 2011 University of Maryland $75K Business Plan Competition ... Winning student and faculty teams were selected from 71 initial ... 6 to teams of expert judges as well as a ...
Cached Biology Technology:Deloitte Seeks Nominations for 2011 Technology Fast 500™ Ranking 2DATATRAK Academy Expands to Include Durham Technical Community College 2DATATRAK Academy Expands to Include Durham Technical Community College 3Mtech Announces Winners of the 2011 University of Maryland $75K Business Plan Competition 2Mtech Announces Winners of the 2011 University of Maryland $75K Business Plan Competition 3Mtech Announces Winners of the 2011 University of Maryland $75K Business Plan Competition 4Mtech Announces Winners of the 2011 University of Maryland $75K Business Plan Competition 5
(Date:7/22/2014)... and engineering professor with an expertise in big data ... grant worth more than $600,000 to develop an interactive ... Huang, an associate professor, will lead the project with ... Science and Engineering department. Scientists are compiling and studying ... of genes involved in fruit fly development are commonly ...
(Date:7/22/2014)... the Monell Center reveals that simply believing that an ... asthmatics for at least 24 hours following exposure. The ... health-related outcomes. , "Asthmatics often are anxious about scents ... harmful, our bodies react as if that odor is ... a Monell physiologist. "Both patients and care providers need ...
(Date:7/22/2014)... not be good for the average steak, but it ... prepare to survive Alaska,s winter cold. , "Alaska wood ... a steak does in your freezer and the frog ... shape than the steak," said Don Larson, University of ... recent paper demonstrating that freeze tolerance in Alaska wood ...
Breaking Biology News(10 mins):UT Arlington big data team wins $600,000 NSF grant to build gene expression database 2In asthma, it's not just what you smell, but what you think you smell 2In asthma, it's not just what you smell, but what you think you smell 3Alaska frogs reach record lows in extreme temperature survival 2Alaska frogs reach record lows in extreme temperature survival 3
... sees the use of terahertz rays as a critical ... terrorist activities. Federici and his research team recently ... in their laboratory that uses a terahertz imaging system. ... to scan airport passengers quickly and efficiently. "Video-Rate terahertz ...
... United States and Europe has identified more than a dozen ... common forms of kidney disease. The team, known as the ... than 65,000 individuals of European descent. Common variations in several ... poor kidney function or chronic kidney disease than in those ...
... than one third of Americans are considered obese and approximately ... (BMI) greater than 40. The April issue of the ... ever-increasing use of bariatric surgery to control the excess morbidity ... that registered dietitians play in the management of patients who ...
Cached Biology News:NJIT physicist sees terahertz imaging as ultimate defense against terrorism 2NJIT physicist sees terahertz imaging as ultimate defense against terrorism 3International research team discovers novel genes influencing kidney disease risk 2Registered dietitians play essential role in management of gastric bypass patients 2Registered dietitians play essential role in management of gastric bypass patients 3
Chicken Anti-Human DNA-directed RNA Polymerase II 7.6 Kd Polypeptide (POLR2L Polyclonal AntibodyOther Sub-Family: not assigned-Other Applications: ELISA, Western Blot...
...
The BD CBA Human Apoptosis Kit can be used to quantitatively measure,cleaved PARP, Bcl-2, and active Caspase-3 protein levels in a single sample. The,Kit performance has been optimized for analysis o...
BD Primaria 35 mm Cell Culture Dish with Easy-Grip design, surface-modified polystyrene for enhanced cell culture. (20/sp, 200/ca) Packaging: 20 / sleeve, 500 / case ...
Biology Products: